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Eukaryotic initiation factor 6 is rate-limiting in translation, growth and transformation

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Abstract

Cell growth and proliferation require coordinated ribosomal biogenesis and translation. Eukaryotic initiation factors (eIFs) control translation at the rate-limiting step of initiation1,2. So far, only two eIFs connect extracellular stimuli to global translation rates3: eIF4E acts in the eIF4F complex and regulates binding of capped messenger RNA to 40S subunits, downstream of growth factors4,5, and eIF2 controls loading of the ternary complex on the 40S subunit and is inhibited on stress stimuli6,7. No eIFs have been found to link extracellular stimuli to the activity of the large 60S ribosomal subunit. eIF6 binds 60S ribosomes precluding ribosome joining in vitro8,9,10. However, studies in yeasts showed that eIF6 is required for ribosome biogenesis rather than translation11,12,13,14. Here we show that mammalian eIF6 is required for efficient initiation of translation, in vivo. eIF6 null embryos are lethal at preimplantation. Heterozygous mice have 50% reduction of eIF6 levels in all tissues, and show reduced mass of hepatic and adipose tissues due to a lower number of cells and to impaired G1/S cell cycle progression. eIF6+/- cells retain sufficient nucleolar eIF6 and normal ribosome biogenesis. The liver of eIF6+/- mice displays an increase of 80S in polysomal profiles, indicating a defect in initiation of translation. Consistently, isolated hepatocytes have impaired insulin-stimulated translation. Heterozygous mouse embryonic fibroblasts recapitulate the organism phenotype and have normal ribosome biogenesis, reduced insulin-stimulated translation, and delayed G1/S phase progression. Furthermore, eIF6+/- cells are resistant to oncogene-induced transformation. Thus, eIF6 is the first eIF associated with the large 60S subunit that regulates translation in response to extracellular signals.

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Figure 1: eIF6 reduction leads to diminished body weight and affects liver growth at the proliferation level.
Figure 2: eIF6 reduction results in 80S complex accumulation and a blunted translational response to insulin.
Figure 3: eIF6 reduction in the cytoplasmic compartment does not affect 60S ribosomal biogenesis.
Figure 4: eIF6 reduction impairs G1/S progression in synchronized cells and causes reduced transformation.

Change history

  • 24 November 2008

    An error was corrected in Fig. 4c on 24 Nov 2008; see PDF for details.

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Acknowledgements

This work was supported by grants AIRC (S.B., P.C.M.), TELETHON GGB05043, CARIPLO 0578 (S.B.) and NIH-RO1 (H.K.). A.B. is supported by grant AICR 05-360. The manuscript has been improved thanks to suggestions from N. Offenhaeuser and A. Boletta. We are indebted to P. G. Pelicci for anti-NPM antibodies, H. Hirai for preliminary soft agar assays, S. Modina for blastocyst preparation, M. Vidali for hepatocytes preparation, M. Malosio for insulin receptor antibodies, S. Gregori for FACS analysis, D. Bartel for reporter constructs, G. Manfioletti for HMGA2 antibodies and F. Loreni for rpS19 antibodies. We acknowledge L. Magri for preliminary experiments, and V. Volta and S. Grosso for suggestions.

Author Contributions V.G., A.M., A.M.B., H.K. and S.B. planned the experiments; V.G., A.M., A.M.B., A.B. and S.B. performed the experiments; all authors analysed the data; and V.G. and S.B. wrote the paper. All authors discussed the results and contributed to the manuscript.

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Correspondence to Stefano Biffo.

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Gandin, V., Miluzio, A., Barbieri, A. et al. Eukaryotic initiation factor 6 is rate-limiting in translation, growth and transformation. Nature 455, 684–688 (2008). https://doi.org/10.1038/nature07267

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